The aim: This study was undertaken to identify antibiotic resistance of Salmonella Typhi in specimens of gall bladder tissue after cholecystectomy. Materials and methods: Salmonella Typhi identification from the isolates have been depended on morphology of the colony and biochemical tests as a first step in identification while final identification has been achieved by the automated VITEK-2 compact system then PCR technique. Results: Depending to finding via the VITEK tests and PCR technique and thirty-five Salmonella Typhi sample have been obtained. This research shown that about 35 (70%) positive result contains, 12 (34.3%) isolates was positive from stool and 23(65.7%) from gall bladder tissue. The results revealed difference in S. Typhi resistance to some antibiotics, where S. Typhi has wide-ranging sensitivity: 35 (100%) to Cefepime, Cefixime and Ciprofloxacin and revealed great sensitivity 22 (62.8%) to Ampicillin. S. Typhi isolates proved extremely resistant 19 (54.2%) and 25 (71.4%) to Trimethoprim/Sulphamethoxazole and Chloramphenicol respectively. Increment in the rate of Salmonella that has multidrug resistance to chloramphenicol, ampicillin, furazolidonecotrimoxazole, streptomycin, and tetracycline is a developing problem and worldwide worry matte. Conclusions: Resistant forms of Salmonella enteric serotype Typhi were detected with increment in the rate of multidrug resistance to chloramphenicol, ampicillin, and tetracycline so currently, Cefepime, Cefixime and Ciprofloxacin and revealed great sensitivity and have become the mainstay of treatment. Challenging difficult which rises in this study is the extend of Multidrug resistant strain (MDR) of S. Typhi.